LOCUS       sequence-84608-e        7926 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7926)
  AUTHORS   Binder A, Lambert J, Morbitzer R, Popp C, Ott T, Lahaye T, Parniske 
            M
  TITLE     A modular plasmid assembly kit for multigene expression, gene 
            silencing and silencing rescue in plants.
  JOURNAL   PLoS One. 2014 Feb 13;9(2):e88218. doi: 
            10.1371/journal.pone.0088218. eCollection 2014.
  PUBMED    24551083
REFERENCE   2  (bases 1 to 7926)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2014-02-13"; volume: "9(2):e88218. doi"; pages: " 
            10.1371/journal.pone.0088218. eCollection 2014"
COMMENT     Sequence Label: LIIIbeta fin
FEATURES             Location/Qualifiers
     source          1..7926
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    18..158
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     73..92
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(173..190)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(344..932)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(424..443)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1002..1021)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1003..1021)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1026..1042)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1026..1042)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1144..1163)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1155..1964
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1222..1241)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    2220..2244
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        join(2423..2428,2429..2446,2447..2453)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2507..3166
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2573..2592)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3508..3813
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     3692..3711
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_feature    3884..3908
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             5209..5838
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             6267..7340
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      7406..7600
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(7836..7858)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact
       61 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
      121 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca
      181 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
      241 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
      301 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
      361 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
      421 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
      481 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
      541 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
      601 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
      661 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
      721 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
      781 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
      841 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
      901 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
      961 ctgacgctca taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt
     1021 acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca
     1081 ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta
     1141 atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca
     1201 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     1261 cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     1321 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     1381 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     1441 ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     1501 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     1561 attgtccttt taacagcgat cgcgtatttc gtcttgctca ggcgcaatca cgaatgaata
     1621 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     1681 tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     1741 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     1801 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     1861 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     1921 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt
     1981 ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa
     2041 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agacccctgc cggtaacatg
     2101 agcaaagtct gccgccttac aacggctctc ccgctgacgc cgtcccggac tgatgggctg
     2161 cctgtatcga gtggtgattt tgtgccgagc tgccggtcgg ggagctgttg gctggctggt
     2221 ggcaggatat attgtggtgt aaacaaattg acgcttagac ggattacgtt gtcttcaaag
     2281 ctgaacgaga aacgtaaaat gatataaata tcaatatatt aaattagatt ttgcataaaa
     2341 aacagactac ataatactgt aaaacacaac atatccagtc actatggcgg cccctcgagg
     2401 ggccgcatta ggcaccccag gctttacact ttatgcttcc ggctcgtata atgtgtggat
     2461 tttgagttag gatccgtcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat
     2521 cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt
     2581 tcagtcagtt gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt
     2641 aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg
     2701 cctgatgaat gctcatccgg aattccgtat ggcaatgaaa gacggtgagc tggtgatatg
     2761 ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct
     2821 ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc
     2881 gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt
     2941 gtcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa
     3001 cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat
     3061 gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct
     3121 taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatcta gaggatccgg
     3181 cttactaaaa gccagataac agtatgcgta tttgcgcgct gatttttgcg gtataagaat
     3241 atatactgat atgtataccc gaagtatgtc aaaaagaggt atgctatgaa gcagcgtatt
     3301 acagtgacag ttgacagcga cagctatcag ttgctcaagg catatatgat gtcaatatct
     3361 ccggtctggt aagcacaacc atgcagaatg aagcccgtcg tctgcgtgcc gaacgctgga
     3421 aagcggaaaa tcaggaaggg atggctgagg tcgcccggtt tattgaaatg aacggctctt
     3481 ttgctgacga gaacaggggc tggtgaaatg cagtttaagg tttacaccta taaaagagag
     3541 agccgttatc gtctgtttgt ggatgtacag agtgatatta ttgacacgcc cgggcgacgg
     3601 atggtgatcc ccctggccag tgcacgtctg ctgtcagata aagtcccccg tgaactttac
     3661 ccggtggtgc atatcgggga tgaaagctgg cgcatgatga ccaccgatat ggccagtgtg
     3721 ccggtttccg ttatcgggga agaagtggct gatctcagcc accgcgaaaa tgacatcaaa
     3781 aacgccatta acctgatgtt ctggggaata taaatgtcag gctcccttat acacagccag
     3841 tctgcaggtc gacgaagact ttctggttta aactatcagt gtttgacagg atatattggc
     3901 gggtaaacct aagagaaaag agcgtttatt agaataatcg gatatttaaa agggcgtgaa
     3961 aaggtttatc cgttcgtcca tttgtatgtg catgccaacc acagggttcc cctcgggatc
     4021 aaagtacttt gatccaaccc ctccgctgct atagtgcagt cggcttctga cgttcagtgc
     4081 agccttcttc tgaaaacgac atgtcgcaca agtcctaagt tacgcgacag gctgccgccc
     4141 tgcccttttc ctggcgtttt cttgtcgcgt gttttagtcg cataaagtag aatacttgcg
     4201 actagaaccg gagacattac gccatgaaca agagcgccgc cgctggcctg ctgggctatg
     4261 cccgcgtcag caccgacgac caggacttga ccaaccaacg ggccgaactg cacgcggccg
     4321 gctgcaccaa gctgttttcc gagaagatca ccggcaccag gcgcgaccgc ccggagctgg
     4381 ccaggatgct tgaccaccta cgccctggcg acgttgtgac agtgaccagg ctagaccgcc
     4441 tggcccgcag cacccgcgac ctactggaca ttgccgagcg catccaggag gccggcgcgg
     4501 gcctgcgtag cctggcagag ccgtgggccg acaccaccac gccggccggc cgcatggtgt
     4561 tgaccgtgtt cgccggcatt gccgagttcg agcgttccct aatcatcgac cgcacccgga
     4621 gcgggcgcga ggccgccaag gcccgaggcg tgaagtttgg cccccgccct accctcaccc
     4681 cggcacagat cgcgcacgcc cgcgagctga tcgaccagga aggccgcacc gtgaaagagg
     4741 cggctgcact gcttggcgtg catcgctcga ccctgtaccg cgcacttgag cgcagcgagg
     4801 aagtgacgcc caccgaggcc aggcggcgcg gtgccttccg tgaggacgca ttgaccgagg
     4861 ccgacgccct ggcggccgcc gagaatgaac gccaagagga acaagcatga aaccgcacca
     4921 ggacggccag gacgaaccgt ttttcattac cgaagagatc gaggcggaga tgatcgcggc
     4981 cgggtacgtg ttcgagccgc ccgcgcacat ctcaaccgtg cggctgcatg aaatcctggc
     5041 cggtttgtct gatgccaagc tggcggcctg gccggccagc ttggccgctg aagaaaccga
     5101 gcgccgccgt ctaaaaaggt gatgtgtatt tgagtaaaac agcttgcgtc atgcggtcgc
     5161 tgcgtatatg atgcgatgag taaataaaca aatacgcaag gggaacgcat gaaggttatc
     5221 gctgtactta accagaaagg cgggtcaggc aagacgacca tcgcaaccca tctagcccgc
     5281 gccctgcaac tcgccggggc cgatgttctg ttagtcgatt ccgatcccca gggcagtgcc
     5341 cgcgattggg cggccgtgcg ggaagatcaa ccgctaaccg ttgtcggcat cgaccgcccg
     5401 acgattgacc gcgacgtgaa ggccatcggc cggcgcgact tcgtagtgat cgacggagcg
     5461 ccccaggcgg cggacttggc tgtgtccgcg atcaaggcag ccgacttcgt gctgattccg
     5521 gtgcagccaa gcccttacga catatgggcc accgccgacc tggtggagct ggttaagcag
     5581 cgcattgagg tcacggatgg aaggctacaa gcggcctttg tcgtgtcgcg ggcgatcaaa
     5641 ggcacgcgca tcggcggtga ggttgccgag gcgctggccg ggtacgagct gcccattctt
     5701 gagtcccgta tcacgcagcg cgtgagctac ccaggcactg ccgccgccgg cacaaccgtt
     5761 cttgaatcag aacccgaggg cgacgctgcc cgcgaggtcc aggcgctggc cgctgaaatt
     5821 aaatcaaaac tcatttgagt taatgaggta aagagaaaat gagcaaaagc acaaacacgc
     5881 taagtgccgg ccgtccgagc gcacgcagca gcaaggctgc aacgttggcc agcctggcag
     5941 acacgccagc catgaagcgg gtcaactttc agttgccggc ggaggatcac accaagctga
     6001 agatgtacgc ggtacgccaa ggcaagacca ttaccgagct gctatctgaa tacatcgcgc
     6061 agctaccaga gtaaatgagc aaatgaataa atgagtagat gaattttagc ggctaaagga
     6121 ggcggcatgg aaaatcaaga acaaccaggc accgacgccg tggaatgccc catgtgtgga
     6181 ggaacgggcg gttggccagg cgtaagcggc tgggttgcct gccggccctg caatggcact
     6241 ggaaccccca agcccgagga atcggcgtga gcggtcgcaa accatccggc ccggtacaaa
     6301 tcggcgcggc gctgggtgat gacctggtgg agaagttgaa ggccgcgcag gccgcccagc
     6361 ggcaacgcat cgaggcagaa gcacgccccg gtgaatcgtg gcaagcggcc gctgatcgaa
     6421 tccgcaaaga atcccggcaa ccgccggcag ccggtgcgcc gtcgattagg aagccgccca
     6481 agggcgacga gcaaccagat tttttcgttc cgatgctcta tgacgtgggc acccgcgata
     6541 gtcgcagcat catggacgtg gccgttttcc gtctgtcgaa gcgtgaccga cgagctggcg
     6601 aggtgatccg ctacgagctt ccagacgggc acgtagaggt ttccgcaggg ccggccggca
     6661 tggccagtgt gtgggattac gacctggtac tgatggcggt ttcccatcta accgaatcca
     6721 tgaaccgata ccgggaaggg aagggagaca agcccggccg cgtgttccgt ccacacgttg
     6781 cggacgtact caagttctgc cggcgagccg atggcggaaa gcagaaagac gacctggtag
     6841 aaacctgcat tcggttaaac accacgcacg ttgccatgca gcgtacgaag aaggccaaga
     6901 acggccgcct ggtgacggta tccgagggtg aagccttgat tagccgctac aagatcgtaa
     6961 agagcgaaac cgggcggccg gagtacatcg agatcgagct agctgattgg atgtaccgcg
     7021 agatcacaga aggcaagaac ccggacgtgc tgacggttca ccccgattac tttttgatcg
     7081 atcccggcat cggccgtttt ctctaccgcc tggcacgccg cgccgcaggc aaggcagaag
     7141 ccagatggtt gttcaagacg atctacgaac gcagtggcag cgccggagag ttcaagaagt
     7201 tctgtttcac cgtgcgcaag ctgatcgggt caaatgacct gccggagtac gatttgaagg
     7261 aggaggcggg gcaggctggc ccgatcctag tcatgcgcta ccgcaacctg atcgagggcg
     7321 aagcatccgc cggttcctaa tgtacggagc agatgctagg gcaaattgcc ctagcagggg
     7381 aaaaaggtcg aaaaggactc tttcctgtgg atagcacgta cattgggaac ccaaagccgt
     7441 acattgggaa ccggaacccg tacattggga acccaaagcc gtacattggg aaccggtcac
     7501 acatgtaagt gactgatata aaagagaaaa aaggcgattt ttccgcctaa aactctttaa
     7561 aacttattaa aactcttaaa acccgcctgg cctgtgcata actgtctggc cagcgcacag
     7621 ccgaagagct gcaaaaagcg cctacccttc ggtcgctgcg ctccctacgc cccgccgctt
     7681 cgcgtcggcc tatcgcggcc gctggccgct caaaaatggc tggcctacgg ccaggcaatc
     7741 taccagggcg cggacaagcc gcgccgtcgc cactcgaccg ccggcgccca catcaaggca
     7801 ccctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     7861 acagtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     7921 gcgggt
//
